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1.
An. bras. dermatol ; 94(2): 198-203, Mar.-Apr. 2019. tab, graf
Article in English | LILACS | ID: biblio-1001146

ABSTRACT

Abstract BACKGROUND: Psoriasis is a systemic inflammatory disorder that involves complex pathogenic interactions between the innate and adaptive immune systems. The most accepted mechanism in the etiopathogenesis of psoriasis is the induction of inflammation with keratinocyte hyperproliferation. Granulysin (GNLY) is a cytolytic antimicrobial peptide (AMP) that is secreted together with granzyme and perforin from the granules of human cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. It has been immunohistochemically proven that the expression of granulysin is increased in lesions of psoriasis. OBJECTIVE: This study aimed to investigate the relationship between psoriasis disease and granulysin gene polymorphisms. METHODS: GNLY rs7908 and rs10180391 polymorphisms were studied by PCR-RFLP in 100 psoriasis patients under treatment in the Dermatology Polyclinic of Bulent Ecevit University. In addition, 100 healthy individuals with similar age and sex distribution were used as a control group. RESULTS: In the control group, GNLY rs7908 CC genotype was significantly higher than in psoriasis patients (P= 0.031; OR= 0.305; Cl= 0.305 (0.121 - 0.773). In our study, the genotype distributions in patients and control groups were GNLY rs7908 (SNP) GG (51%, 37%), GC (41%, 44%), CC (8%, 19%); GNLY rs10180391 (SNP) from the CC (41%, 44%), CT (42%, % 41), TT (17%, 15%). STUDY LIMITATIONS: The study only included Turkish patients. CONCLUSION: Our findings showed that GNLY rs7908 CC genotype and C allele had a protective effect against psoriasis and decreased the disease severity (according to PASI score), whereas rs10180391 SNP did not show any effective role in psoriasis pathogenesis.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Polymorphism, Genetic/genetics , Psoriasis/genetics , Antigens, Differentiation, T-Lymphocyte/genetics , Psoriasis/etiology , Severity of Illness Index , Case-Control Studies , Gene Expression , Protective Agents , Alleles , Genotype
2.
An. bras. dermatol ; 91(5): 642-644, Sept.-Oct. 2016. graf
Article in English | LILACS | ID: biblio-827761

ABSTRACT

Abstract: Sézary syndrome is a primary cutaneous T-cell lymphoma characterized by the triad of erythroderma, lymphadenopathy and circulating atypical cells. The emergence of new molecular targets has enabled the development of drugs such as alemtuzumab, an anti-CD52 monoclonal antibody, which has shown promising results in the treatment of this entity. We report the case of a 70-year-old male with refractory Sézary syndrome in whom treatment with alemtuzumab achieved an 80% skin lesion clearance with complete haematologic and radiologic response. The treatment was discontinued after 4 months due to adverse effects, with the patient showing a sustained response without disease progression after 13 months of follow-up.


Subject(s)
Humans , Male , Aged , Skin Neoplasms/drug therapy , Sezary Syndrome/drug therapy , Antibodies, Monoclonal, Humanized/therapeutic use , Antineoplastic Agents/therapeutic use , Skin Neoplasms/blood , Blood Cell Count , Antigens, Differentiation, T-Lymphocyte/metabolism , Sezary Syndrome/blood , Treatment Outcome , Alemtuzumab
3.
Chinese Journal of Pathology ; (12): 97-101, 2016.
Article in Chinese | WPRIM | ID: wpr-278556

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the diagnostic role of STAT6 immunohistochemistry in solitary fibrous tumors (SFT)/meningeal hemangiopericytomas (HPC).</p><p><b>METHOD</b>Evaluated the expression of STAT6, vimentin, CD34, EMA, PR, S-100, CD56, GFAP and Ki-67 in a cohort of 37 SFT/meningeal HPC, 30 meningiomas and 30 schwannomas by immunohistochemistry staining.</p><p><b>RESULTS</b>All SFT/meningeal HPC demonstrated nuclear positivity for STAT6, and the proportion of positive tumor cells ranged from 60% to 95%, with no significant difference cases.Vimentin was strongly positive in all cases. CD34, EMA and PR positivity was found in 32 cases, 1 case and 4 cases, respectively.S-100 protein, CD56 and GFAP were negative; Ki-67 labeling index was 1%-8%. However, the meningiomas and schwannomas were negative for STAT6.</p><p><b>CONCLUSIONS</b>STAT6 is a relatively specific biomarker for SFT/meningeal HPC, and may be used in the diagnosis and differential diagnosis of SFT/meningeal HPC, especially for the atypical cases, and allows the precise pathologic diagnosis of SFT/meningeal HPC.</p>


Subject(s)
Antigens, CD , Antigens, CD34 , Antigens, Differentiation, T-Lymphocyte , Biomarkers, Tumor , Diagnosis, Differential , Glial Fibrillary Acidic Protein , Hemangiopericytoma , Chemistry , Diagnosis , Humans , Immunohistochemistry , Ki-67 Antigen , Meningeal Neoplasms , Chemistry , Diagnosis , Meningioma , Chemistry , Diagnosis , Neurilemmoma , Chemistry , Diagnosis , S100 Proteins , STAT6 Transcription Factor , Solitary Fibrous Tumors , Chemistry , Diagnosis , Vimentin
4.
J. appl. oral sci ; 23(5): 536-546, Sept.-Oct. 2015. tab, graf
Article in English | LILACS, BBO | ID: lil-764159

ABSTRACT

In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Recently, our research group has reported a differential immunogenicity when T lymphocytes were stimulated with these different serotypes. In particular, it was demonstrated that the serotype b of A. actinomycetemcomitans has a stronger capacity to trigger Th1- and Th17-type cytokine production.Objective This study aimed to quantify the expression of different CC chemokines (CCLs) and receptors (CCRs) in T lymphocytes stimulated with the differentA. actinomycetemcomitans serotypes. In addition, the expression of the transcription factors T-bet, GATA-3, RORC2, and Foxp3, master-switch genes implied in the Th1, Th2, Th17, and T-regulatory differentiation, respectively, was analysed in order to determine T-cell phenotype-specific patterns of CCL and CCR expression upon A. actinomycetemcomitans stimulation.Material and Methods Human naïve CD4+ T lymphocytes were obtained from healthy subjects and stimulated with autologous dendritic cells primed with the differentA. actinomycetemcomitans serotypes. The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR. Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes. In addition, these higher expression levels of CCLs and CCRs positively correlated with the increased levels of T-bet and RORC2 when T lymphocytes were stimulated with the serotype b.Conclusion A T-lymphocyte response biased towards a Th1- and Th17-pattern of CCL and CCR expression was detected under stimulation with the serotype b ofA. actinomycetemcomitans.


Subject(s)
Humans , Male , Female , Adult , Young Adult , Aggregatibacter actinomycetemcomitans/immunology , Antigens, Differentiation, T-Lymphocyte/analysis , Chemokines, CC/analysis , Receptors, CCR/analysis , T-Lymphocytes/immunology , Aggregatibacter actinomycetemcomitans/genetics , Antigens, Differentiation, T-Lymphocyte/genetics , Antigens, Differentiation, T-Lymphocyte/immunology , Cell Differentiation/immunology , Cells, Cultured , Chemokines, CC/genetics , Chemokines, CC/immunology , Dendritic Cells/immunology , Flow Cytometry , Lymphocyte Activation , Polymerase Chain Reaction , Receptors, CCR/genetics , Receptors, CCR/immunology , Serogroup
5.
Article in Chinese | WPRIM | ID: wpr-255161

ABSTRACT

<p><b>OBJECTIVE</b>To construct the expression vector pLCK-CD69-IRES-EGFP that contains mouse cell surface activation protein CD69 and enhanced green fluorescent protein(EGFP),and to generate CD69 transgenic mice based on this vector.</p><p><b>METHODS</b>First, RNA was extracted from mouse lung tissue and cDNA was synthesized via reverse transcription. PCR primer was designed through the PubMed searching, then mouse CD69 DNA fragment was amplified with PCR. Second, this DNA fragment was subcloned to the pInsulater-LCK-IRES-EGFP plasmid and constructed the transgenic vector after the verification of nucleotide sequence. Third, the expression vector was then transfected into 293 T cells and its expression in 293 T cells was observed under fluorescence microscope. Last, microinjection was performed to transfer the expression vector pLCK-CD69-IRES-EGFP into fertilized eggs, which were implanted into pseudo-pregnant recipient mice. After birth the tail samples of the pups were obtained for the purpose of genotyping to determine the transgenic founders. Fluorescence microscope and flow cytometer were used to measure the expression of CD69 on cells.</p><p><b>RESULTS</b>The construction of the expression vector pLCK-CD69-IRES-EGFP was verified by enzyme digestion and DNA sequencing. The transfected 293 T cell showed expression of the protein under fluorescence microscope. Identification of PCR for the tail tissue of the pups confirmed the present of CD69 transgene and resting lymphocytes demonstrated the expression of CD69.</p><p><b>CONCLUSION</b>The construction of expression vector pLCK-CD69-IRES-EGFP and generation of CD69 transgenic mice have been successfully processed, which lays a foundation of the solid pattern studies in inflammatory diseases.</p>


Subject(s)
Animals , Antigens, CD , Genetics , Antigens, Differentiation, T-Lymphocyte , Genetics , DNA, Complementary , Genetic Vectors , Genotype , Green Fluorescent Proteins , Genetics , Lectins, C-Type , Genetics , Mice , Mice, Transgenic , Plasmids , Polymerase Chain Reaction , Sequence Analysis, DNA , Transfection
6.
Acta Pharmaceutica Sinica ; (12): 482-489, 2014.
Article in English | WPRIM | ID: wpr-245058

ABSTRACT

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.


Subject(s)
Animals , Anti-Inflammatory Agents , Pharmacology , Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Arctium , Chemistry , Cell Cycle Checkpoints , Cell Proliferation , Cytokines , Metabolism , Female , Furans , Pharmacology , Interferon-gamma , Metabolism , Interleukin-10 , Metabolism , Interleukin-2 , Metabolism , Interleukin-2 Receptor alpha Subunit , Metabolism , Interleukin-4 , Metabolism , Interleukin-6 , Metabolism , Ionomycin , Pharmacology , Lectins, C-Type , Metabolism , Lignans , Pharmacology , Lymphocyte Activation , Mice , Mice, Inbred BALB C , Plants, Medicinal , Chemistry , T-Lymphocytes , Cell Biology , Allergy and Immunology , Tetradecanoylphorbol Acetate , Pharmacology , Tumor Necrosis Factor-alpha , Metabolism
7.
Hematology, Oncology and Stem Cell Therapy. 2013; 6 (2): 49-57
in English | IMEMR | ID: emr-140985

ABSTRACT

Research has implied that the immune system plays a role in the pathogenesis of MDS and that T-cells are reacting to tumour antigen present on the surface of the malignant cells. This could imply that the immune system could be utilized to generate immune based therapy. The aim of this pilot study was to examine the feasibility of studying this further by analysing the interaction of dendritic cells with T-cells in a small cohort of MDS patients. Dendritic cells were generated in 6 MDS patients and 9 controls by culturing monocytes with GMCSF and IL-4. After activation with LPS and TNFalpha, the dendritic cells were analyzed for expression of co-stimulatory and activation antigens. Thereafter, they were co-cultured with T-cells and the T-cell response was examined by measuring the % change in expression of the activation antigen CD69. MDS MoDC had reduced expression of HLA-DR [p=0.006], CD11c [p=0.0004], CD80 [p=0.03] and CD86 [p=0.003], while resting T-cells from MDS patients had higher expression of the activation antigen CD69 on all subsets. The % change in CD69 expression increased significantly for both the control and MDS T-cells after co-culture with allogeneic dendritic cells, however this change was lower in the MDS group. Despite the increased CD69 expression prior to culture, MDS MoDC significantly up-regulated CD69 expression on autologous T-cells to values that were statistically higher than control cells. This initial study suggests that the T-cells in MDS are able to respond to dendritic cells and are therefore probably not part of the malignant clone. It further implies that the dendritic cell population could be capable of presenting antigen and initiating an immune response and therefore further study is both feasible and warranted


Subject(s)
Humans , Male , Female , Monocytes , T-Lymphocytes , Myelodysplastic Syndromes , Pilot Projects , Antigens, Differentiation, T-Lymphocyte , Antigens, CD , Lectins, C-Type
8.
Article in English | WPRIM | ID: wpr-251440

ABSTRACT

The role of hepatic CD69+ natural killer (NK) cells in virus-induced severe liver injury and subsequent hepatic failure is not well defined. In this study, a mouse model of fulminant liver failure (FHF) induced by murine hepatitis virus strain 3 (MHV-3) was used to study the role of hepatic CD69+NK cells in the development of FHF. The CD69 expression in NK cells in the liver, spleen, bone marrow and peripheral blood was detected by using flow cytometry. The correlation between the CD69 level in hepatic NK cells and liver injury was studied. The functional marker (CD107a), and activating and inhibitory receptor (NKG2D and NKG2A) expressed on CD69+NK cells and CD69-NK cells were detected by using flow cytometry. Pro-inflammatory cytokines (IL-9, IFN-γ and TNF-α) were also examined by using intracellular staining. After MHV-3 infection, the number of CD69+NK cells in the liver of BALB/cJ mice was increased markedly and peaked at 72 h post-infection. Similar changes were also observed in the spleen, bone marrow and peripheral blood. Meanwhile, the CD69 expression in hepatic NK cells was highly correlated with the serum level of ALT and AST. The expression of CD107a and NKG2D, as well as the production of TNF-α, IFN-γ and IL-9 in hepatic CD69+NK cells was all significantly up-regulated during 48-72 h post-infection. In contrast, the NKG2A expression was increased in hepatic CD69-NK cells but not in CD69+NK cells. These results suggested that hepatic CD69+NK cells play a pivotal role in the pathogenesis of FHF by enhancing degranulation and cytotoxic ability of NK cells and increasing the production of pro-inflammatory cytokines.


Subject(s)
Animals , Antigens, CD , Allergy and Immunology , Antigens, Differentiation, T-Lymphocyte , Allergy and Immunology , Coronavirus Infections , Allergy and Immunology , Female , Hepatitis, Viral, Animal , Allergy and Immunology , Killer Cells, Natural , Allergy and Immunology , Lectins, C-Type , Allergy and Immunology , Mice , Mice, Inbred BALB C , Murine hepatitis virus , Allergy and Immunology
9.
Article in Chinese | WPRIM | ID: wpr-232727

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of MHSP65-TCL on melanoma and its effect on the activity of the immunocytes.</p><p><b>METHODS</b>MHSP65-TCL was prepared by mixing MHSP65 with TCL derived from B16 melanoma cell lysate by repeated freezing and thawing. The MHSP65-TCL vaccine was administered in mice bearing B16 melanoma, and the changes in melanoma growth was observed. To investigate the influence of TCL in MHSP65-TCL on the activity of the immunocytes, we co-cultured TCL and mouse spleen cells in vitro, and analyzed CD69 expression on the cells, cell apoptosis, and levels of IL-10 and IFN-γ in the cell culture supernatant.</p><p><b>RESULTS</b>The MHSP65-TCL vaccine showed an anti-melanoma effect in the tumor-bearing mice. In the in vitro experiment, TCL in MHSP65-TCL strongly stimulated the activation of mouse spleen cells while causing apoptosis in some of the immunocytes and promoting cellular IL-10 secretion, but not IFN-γ.</p><p><b>CONCLUSIONS</b>MHSP65-TCL derived from B16 melanoma cells has an anti-melanoma effect mediated by the activation of immunocytes. TCL in MHSP65-TCL also has immunosuppressive effect on immunocytes possibly due to the presence of suppressive components in TCL, and identifying and eliminating these components may potentially improve the anti-tumor actovoty of MSHP65-TCL vaccine.</p>


Subject(s)
Animals , Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Apoptosis , Bacterial Proteins , Allergy and Immunology , Cancer Vaccines , Cell Extracts , Allergy and Immunology , Cell Line, Tumor , Chaperonin 60 , Allergy and Immunology , Female , Interferon-gamma , Metabolism , Interleukin-10 , Metabolism , Lectins, C-Type , Metabolism , Melanoma, Experimental , Allergy and Immunology , Pathology , Mice , Mice, Inbred C57BL , Random Allocation , Spleen , Cell Biology , Allergy and Immunology , Metabolism , Tumor Burden , Allergy and Immunology
10.
IBJ-Iranian Biomedical Journal. 2013; 17 (4): 194-199
in English | IMEMR | ID: emr-148457

ABSTRACT

Rheumatoid arthritis [RA] is a chronic inflammatory disease with many genetic factors predisposing to disease susceptibility. The aim of the present study was to investigate the impact of CD226 rs727088 and rs763361 polymorphisms and susceptibility to RA in a sample of the Iranian population. This case-control study was carried out on 100 patients with RA and 104 healthy subjects. The polymorphisms were determined using tetra amplification refractory mutation system-polymerase chain reaction assay. The rs763361 [Gly307Ser] polymorphism increased the risk of RA in codominant, dominant and recessive-tested inheritance models [odds ratio [OR] = 3.18, 95% confidence intervals [95% CI] = 1.44-7.02, P = 0.004, CC vs. TT, and OR = 1.98, 95% CI = 1.10-3.57, P = 0.023, CC vs. CT-TT, and OR = 2.61, 95% CI = 1.26-5.37, P = 0.010, CC + CT vs. TT, respectively]. In addition, the rs763361 T allele increased the risk of RA [OR = 2.06, 95% CI = 1.38-3.08, P<0.001]. However, no significant difference was observed among the groups regarding CD226 rs727088 polymorphism [Chi[2] = 3.20, P = 0.202]. Our finding showed that CD226 rs763361, but not rs727088, gene polymorphism increased the risk of RA in a sample of the Iranian population


Subject(s)
Humans , Female , Male , Polymorphism, Genetic , Antigens, Differentiation, T-Lymphocyte
11.
Hanyang Medical Reviews ; : 39-44, 2013.
Article in Korean | WPRIM | ID: wpr-199836

ABSTRACT

Asthma is a complex immune mediated chronic inflammatory lung disease characterized by chronic inflammation of the airways, airway hyper-responsiveness and airway obstruction, and the prevalence of this disease has increased in recent years. It is well known that many features of allergic asthma are consequences of Th2 cell dominated immune responses against allergens, thus allergen specific Th2 cells play a critical role in the pathogenesis. In this review, we will discuss the properties of common indoor and outdoor allergens including house dust mite, fungus, pollen and cockroach, the activation and differentiation of naive CD4 T cells by protease allergens, how specific allergens modify host's immune system to mediate immune evasion, and regulation of homing receptor expression and trafficking of allergen specific Th2 cells. Lastly, we will also overview the general course of pathogenesis of allergic asthma and discuss prospects of development of novel immuno-therapies to asthma.


Subject(s)
Airway Obstruction , Allergens , Antigens, Differentiation, T-Lymphocyte , Asthma , Cockroaches , Fungi , Immune Evasion , Immune System , Inflammation , Lung Diseases , Pollen , Prevalence , Pyroglyphidae , Receptors, Lymphocyte Homing , T-Lymphocytes , Th2 Cells
12.
Article in Korean | WPRIM | ID: wpr-11954

ABSTRACT

Collision tumors of the colon are rare. A 54-year-old man was referred to our hospital for the evaluation of hematochezia. Colonoscopy demonstrated the presence of about 3 cm sized mass in the rectosigmoid junction. After surgical resection, the colonic lesion was histologically composed of two discrete lesions: adenocarcinoma in the superficial layer and poorly differentiated neuroendocrine carcinoma in the deeper layer. We report this case of colonic collision tumor (adenocarcinoma and neuroendocrine carcinoma) with a review of the literature.


Subject(s)
Adenocarcinoma/diagnosis , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/metabolism , Carcinoma, Neuroendocrine/diagnosis , Colonic Neoplasms/diagnosis , Colonoscopy , Humans , Male , Middle Aged , Positron Emission Tomography Computed Tomography , Synaptophysin/metabolism , Tomography, X-Ray Computed
13.
Article in English | WPRIM | ID: wpr-820104

ABSTRACT

OBJECTIVE@#To provide further evidence for the ethnomedicinal use of the Eastern Nigeria mistletoe, Loranthus micranthus (L. micranthus), as an immunostimulant.@*METHODS@#Solvent fractions from the crude extract of the mistletoe plant was obtained and screened by the cell mediated delayed type hypersensitivity reaction (DTHR) model in mice. Then the immunomodulatory potentials of a major lupane triterpenoid ester isolated from an active hexane fraction of the Eastern Nigeria mistletoe was investigated. Three lupeol-based triterpenoid esters: 7β 15α-dihydroxyl-lup-20(29)-ene-3β-palmitate (I), 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-stearate (II) and 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-decadecanoate (III) were isolated from the plant leaves epiphyting on a local kola nut tree and were characterized. Compound 1 was subjected to cell proliferation studies using C57Bl/6 splenocytes at three dose levels (5, 25 and 100 μg/mL) in presence of controls. Furthermore, the effect of this compound on IL-8 receptor expression was evaluated at three doses (1, 5 and 10 μg/mL) using the real time polymerase chain reaction assay.@*RESULTS@#This triterpenoid ester produced some enhancement of the splenocytes at the tested doses but at doses higher than 5 μg/mL caused inhibition of the IL-8 receptor expression.@*CONCLUSIONS@#The present findings support the ethnomedicinal use of the Eastern Nigeria Mistletoe in the management of diseases affecting the immune system. The triterpenoid(s) have some immunomodulatory abilities on splenocytes and IL-8 receptors and may partly account for the overall immunomodulatory activity of this plant.


Subject(s)
Animals , Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Esters , Female , Hypersensitivity, Delayed , Allergy and Immunology , Metabolism , Immunologic Factors , Pharmacology , Lectins, C-Type , Metabolism , Loranthaceae , Chemistry , Lymphocyte Activation , Male , Mice , Mice, Inbred C57BL , Plant Extracts , Pharmacology , Plant Leaves , Chemistry , Rats , Receptors, Interleukin-8 , Spleen , Allergy and Immunology , T-Lymphocytes , Allergy and Immunology , Triterpenes , Chemistry , Pharmacology
14.
Chinese Journal of Hepatology ; (12): 96-100, 2010.
Article in Chinese | WPRIM | ID: wpr-247585

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression profile of immune effector molecules in peripheral natural killer cells (NK) in patients with chronic hepatitis virus B.</p><p><b>METHODS</b>According to the infection status, patients were divided into four experiment groups: normal hepatic function and high HBV DNA level group, normal hepatic function and low HBV DNA level group, abnormal hepatic function and high HBV DNA level group and abnormal hepatic function and low HBV DNA level group. The expression of perforin (PF), granzyme B (Gr B), granulysin (GNLY), tumor necrosis factor alpha (TNFa) and interferon gamma (IFNr) in NK cells were detected by flow cytometer.</p><p><b>RESULTS</b>Compared with control group (31.50%+/-27.64%), the expression of GNLY was significantly increased in normal hepatic function and high HBV DNA level group (59.74%+/-30.82%) and normal hepatic function and low HBV DNA level group (61.89%+/-33.30%); the expression of IFNr in normal hepatic function and high HBV DNA level group (39.89%+/-21.30%) and abnormal hepatic function and high HBV DNA level group (37.54%+/-18.79%) was lower than that in normal control group (57.38%+/-23.69%); the expression of PF, GrB, GNLY in abnormal hepatic function and high HBV DNA level group (35.47%+/-29.64%, 66.55%+/-22.92%, 42.03%+/-33.17%) was lower than that in normal hepatic function and high HBV DNA level group (56.98%+/-38.34%, 81.53%+/-19.58%, 59.74%+/-30.82%) and normal hepatic function and low HBV DNA level groups (62.95%+/-31.98%, 84.51%+/-14.57%, 61.89%+/-33.3%); there were positive correlations between ef PF, Gr B, GNLY, TNFa, and IFNr.</p><p><b>CONCLUSION</b>The expression of IFNr in NK cells from patients with high HBV DNA replication level is lower than that in normal control group; the expression of PF, Gr B and GNLY in NK cells from patients with normal hepatic function is higher than that in NK cells from patients with abnormal hepatic function.</p>


Subject(s)
Adolescent , Adult , Antigens, Differentiation, T-Lymphocyte , Metabolism , Case-Control Studies , Cytokines , Metabolism , DNA, Viral , Blood , Female , Flow Cytometry , Gene Expression Profiling , Granzymes , Metabolism , Hepatitis B virus , Genetics , Allergy and Immunology , Hepatitis B, Chronic , Allergy and Immunology , Pathology , Humans , Killer Cells, Natural , Allergy and Immunology , Metabolism , Liver Function Tests , Male , Middle Aged , Perforin , Metabolism , Virus Replication , Young Adult
15.
Chinese Journal of Pathology ; (12): 19-24, 2010.
Article in Chinese | WPRIM | ID: wpr-273429

ABSTRACT

<p><b>OBJECTIVE</b>To study the expression and localization of co-stimulators in the mucosa of patients with ulcerative colitis (UC), and to explore its role in the pathogenesis of UC.</p><p><b>METHODS</b>Expression of co-stimulators CD86 and inducible co-stimulator (ICOS) was studied by immunohistochemistry on paraffin-embedded mucosal tissue from patients with active UC (64 cases), inactive UC (51 cases) and normal controls (20 cases). Immunostaining for CD28 was also carried out on frozen fresh mucosal tissue sampled from patients with active UC (7 cases), inactive UC (2 cases) and normal controls (5 cases). In addition, expression of CD4, CD8 and CD20 were also examined.</p><p><b>RESULTS</b>In active UC, increased expression of CD86 was not only observed in lamina propria mononuclear cells but also in the intestinal epithelial cells, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in lamina propria mononuclear cells was detected in active UC, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in intestinal epithelial cells was also seen in active UC, as compared with that of inactive UC (P < 0.01). The expression of CD86 was higher in inactive UC than in the normal controls (P < 0.05 or P < 0.01). However, the expression of ICOS showed no statistically significant difference between inactive UC and normal controls. Increased expression of CD28 in active UC, compared with that in inactive UC and normal controls, was also noticed (P < 0.05 or P < 0.01). The number of CD4 or CD8-positive intraepithelial lymphocytes and lymphocytes infiltrating in the lamina propria and small vessel walls was much higher in active UC than in inactive UC and normal controls (P < 0.01). Moreover, the ratio of CD4/CD8 was highest in active UC (P < 0.01). The number of CD20-positive B lymphocytes in lamina propria was also higher in active UC than in inactive UC and normal controls (P < 0.01).</p><p><b>CONCLUSIONS</b>In active UC, CD86 and ICOS were over-expressed in the intestinal epithelial cells and lamina propria mononuclear cells. The phenomenon suggests that abnormal expression of co-stimulators may contribute to the deregulation of acquired immune responses in UC.</p>


Subject(s)
Adult , Aged , Antigens, Differentiation, T-Lymphocyte , Metabolism , B7-2 Antigen , Metabolism , CD28 Antigens , Metabolism , CD4-CD8 Ratio , Case-Control Studies , Colitis, Ulcerative , Metabolism , Pathology , Epithelial Cells , Metabolism , Pathology , Female , Humans , Immunohistochemistry , Inducible T-Cell Co-Stimulator Protein , Intestinal Mucosa , Metabolism , Pathology , Leukocytes, Mononuclear , Metabolism , Pathology , Male , Middle Aged , Mucous Membrane , Metabolism , Pathology , Young Adult
16.
Article in Chinese | WPRIM | ID: wpr-333997

ABSTRACT

Inducible costimulatory molecule (ICOS), a CD28 family member expressed on activated T cells, plays an important roles in T cell activation and effector function. This study was purposed to investigate the effect of blocking ICOS-B7h signal pathway by ICOS-Ig fusion protein on allogeneic reactive T cells and its mechanism. CHO cells stably and highly expressing ICOS-Ig were established, while the human ICOS-Ig fusion protein was harvested and purified from supernatant of CHO cells transfected with pSecTag2/Hygro A-ICOS-Ig. The CD4(+) cells from spleen of C57BL/6 mice were used as reactive cells, the bone marrow derived dendritic cells (DCs) from BALB/C mice were used as stimulatory cells, these cells were treated with different concentrations of ICOS-Ig or human Ig (h-Ig) as control. The results showed that the target protein with molecular weigh 54 kD and endotoxin level < 10 EU/ml was gained. The ICOS-Ig (> or = 10 microg/ml) could significantly inhibited the proliferative effect of allogeneic reactive T cells resulting from stimulation of DCs (p < 0.01). ICOS-Ig did not influence the activation of CD4(+) T cells. ICOS-Ig concentration positively related to the apoptosis of CD4(+) T cells. The percentages of CD4(+) Annexin V(+)PI(-) cells in simple stimulated group, ICOS-Ig 10 microg/ml group and ICOS-Ig 20 microg/ml group were 15.1%, 26.4% and 33.6% respectively. ICOS-Ig decreased secretion of TNFalpha and increased secretion of IL-4. It is concluded that the ICOS-Ig fusion protein has bioactivity of inhibiting T cell proliferation and altering the polarization of T helper cells to Th2 cells which promotes the apoptosis of allogeneic reactive T cells but had no effect on the activation of allo-reactive CD4(+) T cells.


Subject(s)
Animals , Antigens, Differentiation, T-Lymphocyte , Pharmacology , Apoptosis , CD4-Positive T-Lymphocytes , Allergy and Immunology , Metabolism , CHO Cells , Cell Proliferation , Cricetinae , Cricetulus , Inducible T-Cell Co-Stimulator Protein , Interleukin-4 , Bodily Secretions , Lymphocyte Activation , Allergy and Immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Recombinant Fusion Proteins , Pharmacology , Signal Transduction , Th1 Cells , Allergy and Immunology , Metabolism , Th2 Cells , Allergy and Immunology , Metabolism
17.
Chinese Journal of Biotechnology ; (12): 235-241, 2009.
Article in Chinese | WPRIM | ID: wpr-302830

ABSTRACT

The G13 domain derived from granulysin shows high antimicrobial activities against Gram-positive and Gram-negative bacteria but does not lyse Jurkat cells or liposomes. To explore a new approach for high expression of the G13 domain, we fused the sequence encoding G13 to thioredoxin (Trx) gene to construct the recombinant expression vector (pThioHisA-G13). A cyanogen bromide (CNBr) cleavage site was introduced between the Trx and G13 to facilitate final release of the recombinant G13. The recombinant expression vector, pThioHisA-G13, was transformed into E. coli BL21 (DE3). Upon induction by IPTG Trx-G13 fusion protein was expressed and took the form of inclusion bodies counting 58% (W/W) of total cellular proteins. The inclusion body was solved by urea (8 mol/L) and then cleaved by CNBr. We purified the recombinant peptide G13 by one-step cation exchange chromatography. Results of agarose diffuse assay analysis indicated that the recombinant G13 exhibited antibacterial activity. The procedure described in this study will provide a reliable and simple method for highly efficient production of some cationic antimicrobial peptides.


Subject(s)
Anti-Infective Agents , Metabolism , Antigens, Differentiation, T-Lymphocyte , Genetics , Cyanogen Bromide , Pharmacology , Escherichia coli , Genetics , Metabolism , GTP-Binding Protein alpha Subunits, G12-G13 , Genetics , Inclusion Bodies , Metabolism , Protein Structure, Tertiary , Genetics , Recombinant Fusion Proteins , Genetics , Thioredoxins , Genetics , Transfection
18.
Journal of Experimental Hematology ; (6): 1301-1306, 2009.
Article in Chinese | WPRIM | ID: wpr-343298

ABSTRACT

This study was purposed to explore the effect of bone marrow derived mesenchymal stem cells (MSCs) on the expression of CD69 on cytokine-induced killer (CIK)/natural killer (NK) cells derived from cord blood and on the quantity ratio of CD4+CD25+ T regulatory cells in CIK/NK cell culture system using Transwell non-contact cell culture system. The experiments were divided into two groups: Transwell non-contact culture and mixture culture. The ratio of MSC to CIK/ NK cells was 1:20, 1:50 and 1:100. In mixture culture groups, MSC and CIK/NK cells were co-cultured by together contact as the same ratio of Transwell non-contact culture groups. The expression of CD69 on CIK/NK cells, as well as the quantity ratio of CD4+CD25+ T regulatory cells in CIK/NK cell culture were evaluated by flow cytometry. The results showed that the expression of CD69 on CIK/NK cells in experimental groups were significantly lower than that in control group (p<0.001). As to Transwell groups, CD69 expression on the CIK/NK cells at 1:20 ratio of MSC and CIK/NK was significantly lower than that at 1:50 and 1:100 ratio. There were no differences in the expression of CD69 on CIK cells in mixture groups with various MSC ratios, whereas the expression of CD69 on NK cells at 1:20 ratio was significantly lower than that at 1:50 and 1:100. The quantity ratio of CD4+CD25+ cells in CIK/NK cell culture system of experimental groups with MSC co-culture was significantly higher than that in control. As to Transwell groups, the ratio of CD4+CD25+ cells in CIK/NK cell culture system at 1:20 and 1:50 was significantly higher than that at 1:100. The quantity ratio of CD4+CD25+ cells in CIK/NK cell culture system showed significant differences in various mixture groups. As to 1:20 ratio the amount of CD4+CD25+ cells in CIK/NK cell culture system of mixture groups was significantly higher than that in Transwell groups, while there were no differences of the quantity ratio of CD4+CD25+ cells in CIK/NK cell culture at 1:50 and 1:100. It is concluded that either by non-contact Transwell or mixed co-culture, the MSC can suppress the activation of allogeneic CB-CIK/NK cells, which maybe relate to up-regulating the ratio of CD4+CD25+ T regulatory cells in CIK/NK cell culture system in dose-dependent manner.


Subject(s)
Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Cell Culture Techniques , Cells, Cultured , Cytokine-Induced Killer Cells , Allergy and Immunology , Metabolism , Fetal Blood , Cell Biology , Allergy and Immunology , Humans , Lectins, C-Type , Metabolism , Mesenchymal Stem Cells , Cell Biology , T-Lymphocytes, Regulatory , Cell Biology , Metabolism
19.
Acta Pharmaceutica Sinica ; (12): 480-485, 2009.
Article in English | WPRIM | ID: wpr-278235

ABSTRACT

This study investigated the effect of phloretin (Ph) on the proliferation, activation, and cell-cycle distribution of mouse T lymphocytes and NO production and phagocytosis of macrophages. Carboxyfluorescein diacetatesuccinimidyl ester (CFDA-SE) staining plus flow cytometry assay was employed to obtain the proliferation-related index (PI) of lymphocytes. The expression levels of CD69 and CD25 on T lymphocytes stimulated with Con A were evaluated with flow cytometry after staining with fluorescent monoclonal antibody. Cell-cycle distribution of T lymphocytes was analyzed by propidium iodide staining. Griess kit was used to evaluate the NO production and fluorescent microbeads were used to analyze the phagocytosis ability of macrophages. Our results showed that phloretin (40, 60, and 80 micromol x L(-7)) significantly inhibited the proliferation of T lymphocytes and the PI reduced from 1.41 +/- 0.13 to 1.34 +/- 0.16, 1.19 +/- 0.12 and 1.07 +/- 0.06, respectively. Phloretin significantly inhibited the expression of CD69 and CD25 (P < 0.01). The cell cycle distribution analysis showed that phloretin could induce a cell cycle arrest at G0/G1 phase. NO production of LPS +IFN-gamma group of macrophages was (26.72 +/- 3.57) micromol x L(-1), and was significantly reduced by phloretin (P < 0.01). And phagocytosis rate of macrophages was significantly reduced by phloretin (P < 0.01). The results demonstrate that phloretin might be developed into a new immuosuppressive drug.


Subject(s)
Animals , Anti-Inflammatory Agents , Pharmacology , Antigens, CD , Metabolism , Antigens, Differentiation, T-Lymphocyte , Metabolism , Cell Cycle , Cell Proliferation , Female , Immunosuppressive Agents , Pharmacology , Interleukin-2 Receptor alpha Subunit , Metabolism , Lectins, C-Type , Metabolism , Macrophages , Physiology , Bodily Secretions , Mice , Mice, Inbred BALB C , Nitric Oxide , Bodily Secretions , Phagocytosis , Phloretin , Pharmacology , T-Lymphocytes , Cell Biology , Allergy and Immunology
20.
Bauru; s.n; 2009. 128 p. ilus, tab, graf.
Thesis in Portuguese | LILACS, BBO | ID: lil-557724

ABSTRACT

Programmed death-1 (PD-1) é uma proteína de membrana que funciona como um importante regulador negativo da atividade de linfócitos ativa dos, participando, desta forma, do delicado balanço entre ativação e tolerância de células T. Dados recentes têm evidenciado que os mecanismos de tolerância periférica, mediados pela interação PD-1/PD- L1, também impedem uma resposta imune antitumoral eficaz, mesmo em condições nas quais os antígenos tumorais possam ser reconhecidos. Apesar de crescentes evidências demonstrarem o papel da via PD-1 no escape tumoral, pouco se sabe a respeito do seu significado em tumores da cavidade oral. Sabe-se menos ainda sobre a expressão desta molécula em lesões orais pré-neoplásicas. Baseado no exposto, o presente estudo analisou a expressão de PD-1 e seu ligante PD-L1 em lesões de queilite actínica e carcinoma espinocelular de boca através de citometria de fluxo e imunomarcação in situo Os resultados obtidos demonstraram que as células isoladas do sangue periférico e de lesões de queilite actínica e carcinoma espinocelular oral apresentam expressão aumentada de PD-1. A expressão de PD-L1 é mais restrita em queilite actínica, porém é intensa em carcinoma espinocelular oral.


Programmed death-1 (PD-l) is a transmembrane protein that acts as a negative regulator in effector T cells, modulating the delicate balance between effective antimicrobial immune defenses and immune-mediated tissue damage. However, recent evidences suggest that the PD-l: PD-L1 pathway can also block antitumor immune responses even when tumor antigens can be recognized. In spite of growing data indicating the involvement of PD-l in tumor escape, little is known about its role in tumors of oral cavity. In addition, the involvement of PD-l in pre-malignant lesions is an important issue to be clarified. In the present work we investigated the expression of PD-l and PD-L1 in blood and biopsies of patients with actinic cheilitis and oral squamous cell carcinoma by flow cytometry, immunofluorescence and immunohistochemistry. Our data showed that Iymphocytes obtained from peripheral blood and lesion sites exhibited high expression of PD-l in both groups studied Moreover, PD-L1 expression was intense in oral squamous cell carcinoma and moderate in actinic cheilitis.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged, 80 and over , Antigens, Differentiation, T-Lymphocyte , Carcinoma, Squamous Cell/immunology , Mouth Neoplasms/immunology , Cheilitis/immunology , Antigenic Modulation , Cell Separation , Carcinoma, Squamous Cell/etiology , Flow Cytometry , T-Lymphocytes/immunology , Microscopy, Electron, Scanning , Cheilitis/etiology
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